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Enzymes play a pivotal role in biofuels production at competitive price. Biofuels are considered the cornerstone of advance biorefinery development. Bioethanol, biodiesel and biogas are major liquid transportation biofuels which are sustainable, clean and carbon neutral alternatives of conventional petroleum derived fuels. While first generation (1G) ethanol production is well established process, cellulosic ethanol production is still evolving to meet the right economics at commercial scale. Ethanol produced from lignocellulosic biomass so called second-generation (2G) is a cumbersome process and costlier than IG ethanol. Given that high ethanol demand in various sectors, harnessing the lignocellulosic biomass for ethanol production is necessary. Currently, the cost of production and the technical immaturity of the process are major impeding factors in commercialization of 2G ethanol. Cellulases hydrolyse the second-generation biomass into renewable sugars which are fermented into ethanol, biobutanol, and other biofuels. Biodiesel is another promising sustainable fuel made from waste oils, animal fats, vegetal oils and is totally renewable and non-toxic liquid fuel. Lipase catalyse triacylglycerides transesterification reactions making biodiesel. Biogas is a potential energy source which is produced by the anaerobic digestion (AD) of organic wastes in the form of methane. Hydrolases enzymes such as cellulase, hemicellulase, pectinase, ligninase have a crucial role in AD of organic wastes. Fungi are the major producers of cellulases, lipases, ligninases growing on waste feedstocks under submerged or solid-state fermentation process. This talk will present the technological routes of enzymes production, mechanistic action of enzymes, commercial aspects and the crucial role of enzymes in biofuels production. Also, the cutting-edge advancements made in biorefinery will be appraised.